Name: Amplite® Fluorimetric Peroxidase (HRP) Assay Kit *Near Infrared Fluorescence*
Brand: AAT Bioquest
SKU: 11553
Price: 355 USD
Availability: InStock

Amplite® Fluorimetric Peroxidase (HRP) Assay Kit

Near Infrared Fluorescence

Amplite® Fluorimetric Peroxidase (HRP) Assay Kit *Near Infrared Fluorescence* is a fluorimetric assay kit for detection of horseradish peroxidase activity in a microplate format.

Detection Principle: Measures HRP activity using a fluorogenic peroxidase substrate.
Signal Advantage: Near-infrared fluorescence minimizes background from biological sample autofluorescence and autoabsorption. AAT Bioquest
Workflow: One-step, homogeneous, no-wash assay compatible with HTS liquid handling.
Instrument Compatibility: Fluorescence detection at Ex/Em 640/680 nm; absorbance can also be read at 647 ± 5 nm.

HRP dose response was measured with Amplite® Fluorimetric Peroxidase Assay Kit in a solid black 384-well plate using a Gemini fluorescence microplate reader (Molecular Devices).

Protocol

SDS

Catalog	Size	Price	Quantity
11553	500 Tests	Price

Overview

Peroxidase is a small molecule (MW ~40 KD) that can usually be conjugated to an antibody in a 4:1 ratio. Due to its small size, it rarely causes steric hindrance problem with antibody/antigen complex formation. Peroxidase is inexpensive compared to other labeling enzymes. The major disadvantage associated with peroxidase is their low tolerance to many preservatives such as sodium azide that inactivates peroxidase activity even at low concentration. HRP conjugates are extensively used as secondary detection reagents in ELISAs, immuno-histochemical techniques and Northern, Southern and Western blot analyses. We offer this quick (10 min) HRP assay in a one-step, homogeneous, no wash assay system. This kit uses Amplite® IR, our near infrared flurogenic HRP substrate. Amplite® IR generates a substance that has maximum absorption of 647 nm with maximum emission at 670 nm. This near infrared absorption and fluorescence minimize the assay background that is often caused by the autoabsorption and/or autofluorescence of biological samples that rarely absorb light beyond 600 nm. The kit can be used for ELISAs, characterizing kinetics of enzyme reaction and high throughput screening of oxidase inhibitors, etc. The kit provides an optimized 'mix and read' assay protocol that is compatible with HTS liquid handling instruments.

References

View all 49 references: Citation Explorer

Horseradish peroxidase-driven fluorescent labeling of nanotubes with quantum dots

Authors:

Didenko VV, Baskin DS.

Journal:

Biotechniques (2006): 295

Enzymatic oxidation of dipyridamole in homogeneous and micellar solutions in the horseradish peroxidase-hydrogen peroxide system

Authors:

Almeida LE, Imasato H, Tabak M.

Journal:

Biochim Biophys Acta (2006): 216

Recent advances in catalytic peroxidase histochemistry

Authors:

Krieg R, Halbhuber KJ.

Journal:

Cell Mol Biol (Noisy-le-grand) (2003): 547

Vesicular transport route of horseradish C1a peroxidase is regulated by N- and C-terminal propeptides in tobacco cells

Authors:

Matsui T, Nakayama H, Yoshida K, Shinmyo A.

Journal:

Appl Microbiol Biotechnol (2003): 517

Synthesis and purification of horseradish peroxidase-labeled oligonucleotides for tyramide-based fluorescence in situ hybridization

Authors:

van Gijlswijk RP, van de Corput MP, Bezrookove V, Wiegant J, Tanke HJ, Raap AK.

Journal:

Histochem Cell Biol (2000): 175

Spectral properties

Excitation (nm)	648
Emission (nm)	668

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Absorbance microplate reader
Absorbance	647 ± 5 nm
Recommended plate	Clear bottom

Fluorescence microplate reader
Excitation	640nm
Emission	680 nm
Cutoff	665 nm
Recommended plate	Solid black

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Page updated on October 8, 2026

Component A: Amplite® IR Peroxidase Substrate	1 vial
Component B: H2O2	1 vial (3% stabilized solution, 200 μL)
Component C: Assay Buffer	1 bottle (100 mL)
Component D: Horseradish Peroxidase	1 vial (20 units)
Component E: DMSO	1 vial (0.5 mL)